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Insulin/IGF-1 Signaling (IIS) is known to constrain longevity by inhibiting the transcription factor FOXO. How phosphorylation mediated by IIS kinases regulates lifespan beyond FOXO remains unclear. Here, we profile IIS-dependent phosphorylation changes in a large-scale quantitative phosphoproteomic analysis of wild-type and three IIS mutant Caenorhabditis elegans strains. We quantify more than 15,000 phosphosites and find that 476 of these are differentially phosphorylated in the long-lived daf-2/insulin receptor mutant. We develop a machine learning-based method to prioritize 25 potential lifespan-related phosphosites. We perform validations to show that AKT-1 pT492 inhibits DAF-16/FOXO and compensates the loss of daf-2 function, that EIF-2α pS49 potently inhibits protein synthesis and daf-2 longevity, and that reduced phosphorylation of multiple germline proteins apparently transmits reduced DAF-2 signaling to the soma. In addition, an analysis of kinases with enriched substrates detects that casein kinase 2 (CK2) subunits negatively regulate lifespan. Our study reveals detailed functional insights into longevity., How phosphorylation mediated by Insulin/IGF-1 Signaling kinases regulates lifespan remains unclear. Here the authors perform a large-scale quantitative phosphoproteomic analysis of wildtype and IIS mutant C. elegans strains to reveal detailed functional insights into longevity.

Background Ticks have been reported across China, but information on ticks from Chongming Eco-Island of Shanghai is lacking. Understanding the distribution and genetic evolution of ticks on the island is crucial for tick-borne infection control. Methods Ticks were collected from eight towns and four protected areas of Chongming Island during April 2021-March 2022 and were morphologically identified under microscopy. Molecular characterization of the ticks was performed by PCR amplification and sequencing of 12S rRNA and co1 mitochondrial genes. Homology analyses with published sequences obtained from GenBank were conducted for genetic relationships based on neighbor-joining (NJ) analysis of the 12S rRNA and co1 gene loci. To assess genetic evolution, intraspecific and interspecific genetic distances and barcoding gap were assessed based on NJ-K2P analysis of the 12S rRNA locus. Furthermore, genetic diversity and neutrality tests were performed on 12S rRNA sequences. Results A total of 1417/1933 ticks were successfully amplified by PCR of the 12S rRNA and co1 loci. Sequencing and homology analyses revealed four species of ticks, including Haemaphysalis flava (1376/1417 [97.11%], Ha. longicornis (37/1417 [2.61%]), Ha. doenitzi (3/1417 [0.21%]), and Ha. japonica (1/1417 [0.07%]). The intraspecies identity was 100% for Ha. longicornis, 99.11% for doenitzi, and 96.17%, while Ha. japonica had 83.48% intraspecies identity. Phylogenetic analyses showed species specificity, which clustered with homologous sequences. The average K2P distances between and within the species were 0.1351 (range 0.0723–0.1960) and 0.0145 (range 0-0.0946), respectively. The maximum K2P distance within species was in Ha. japonica (0.0946), while the maximum interspecific K2P distance was 0.1960 between Ha. hystricis and Ha. japonica. Genetic diversity indices and neutrality tests revealed that Fu and Li’s D and Tajima’s D values were negative for all species (mean values − 0.9360 and − 1.0310, respectively), suggesting evidence of recent population expansion. Conclusions This study is the first to report the genetic characterization of ticks from Chongming Eco-Island of Shanghai. We identified four species of Haemaphysalis ticks that were genetically similar to other congeners reported in China or worldwide. The knowledge gained from this study serves as a basis to support intervention deployment for effective control of tick-borne infections.

The disease-gene-drug multi-level network constructed by network pharmacology can predict drug targets and has been widely used in the study of material basis and mechanism of action of Chinese medicinal prescriptions. However, most of the current studies have normalized the efficacies of Chinese herbal medicines in the compounds during the construction of the network. There is also a lack of in-depth exploration of the mechanism of synergy among multiple components. This study proposed a network module partition method based on group collaboration and the pharmacological network was weighed according to the traditional Chinese medicine(TCM) theory ofquot;monarch, minister, assistant and guidequot;. Taking the Tanyu Tongzhi Prescription as an example, we constructed its pharmacological network for the treatment of myocardial ischemia-reperfusion injury. The group collaboration module in the network was identified and the network changes before and after the weighting were compared based on the network topology analysis to explore a new method to find the core nodes of the network as well as the core drugs that affected the efficacy of the compounds. The results showed that the module partition method based on group collaboration could be used to identify and partition group collaboration mo-dules in pharmacological networks of compounds. The proposed weighted network based on the TCM theory ofquot;monarch, minister, assistant, and guidequot; could identify and partition the modules based on the characteristics of the pharmacological network. The identification and partition results of modules of Tanyu Tongzhi Prescription in the weighted network were superior to those in the unweighted network. The weighted closeness centrality(WCC) evaluation method was conducive to finding key nodes and relations in the network as compared with traditional methods, thereby providing a basis for analyzing the core components of drugs and extracting more accurate drug components and targets.

The widespread application of quaternary ammonium compounds (QAC) has posed a serious hazard to the environment and human being, and high concentration of Cl

This paper explores the impact of joining centralized drug procurement of China on the profitability of medical enterprises by the difference-in-difference (DID) model. When centralized procurement cannot bring enough cost savings to enterprises, the price competition caused by centralized procurement will lead to the decline of enterprise profits. In the short term, the negative impact of China's drug centralized procurement policy on the net profit of enterprises is not obvious in the year when enterprises win the bid. After the government officially purchases from pharmaceutical enterprises, the negative impact of the drug centralized procurement policy of China on the net profit of enterprises begins to appear gradually. Therefore, the generic drug manufacturers increase R&D investment and have their own heavy products of original drugs as soon as possible to enhance their core competitiveness.

Chemical structure of azo dyes molecules showed significant influence on their decolorization rate, while the structure-activity relationship between chemical structure and their reduction decolorization rate is not fully understand. In this study, we found that azo dye molecule with closer position for electron-withdrawing substituent to azo bond resulted in faster chemical and biotic reduction rate with or without presence of carbon nanotubes (CNTs), while electron-repulsive substituent closer to azo bond leading to slower azo dye chemical and biotic reduction rate no matter with or without presence of CNTs. Additionally, galvanic cell experiments implied that electron transfer process may play important roles for both chemical and biological reduction decolorization of azo dyes, and CV results indicated that the higher (azo bond breakage) reduction wave potential corresponding to a faster azo dye chemical decolorization reaction. Finally, the results of Lowest Unoccupied Molecular Orbital (LUMO) energy established that lower LUMO energy for azo dye corresponding to a faster chemical decolorization reaction. This study not only offer systematized relationships between structure property of azo dye and their decolorization rate, but also provide a universal and propagable reduction rules.

High alcohol-producing Klebsiella pneumoniae (HiAlc Kpn) in the gut microbiota had been demonstrated to be the causative agent of fatty liver disease (FLD). However, the catabolic pathways for alcohol production in vivo remain unclear. Here, we characterized the genome of HiAlc and medium alcohol-producing (MedAlc) Kpn and constructed an adh (an essential gene encoding alcohol dehydrogenase) knock-out HiAlc Kpn W14 strain (W14Δadh) using CRISPR-Cas9 system. Subsequently, we established the mouse model via gavage administration of HiAlc Kpn W14 and W14 Δadh strains, respectively. Proteome and metabolome analysis showed that 10 proteins and six major metabolites involved in the 2,3-butanediol fermentation pathway exhibited at least a three-fold change or greater during intestinal growth. Compared with HiAlc Kpn W14-fed mice, W14Δadh-fed mice with weak alcohol-producing ability did not show apparent pathological changes at 4 weeks, although some steatotic hepatocytes were observed at 12 weeks. Our data demonstrated that carbohydrate substances are catabolized to produce alcohol and 2,3-butanediol via the 2,3-butanediol fermentation pathway in HiAlc Kpn, which could be a promising clinical diagnostic marker. The production of high amounts of endogenous alcohol is responsible for the observed steatosis effects in hepatocytes in vivo.

Pyrogenic carbonaceous matter (PCM) catalyzes the transformation of a range of organic pollutants by sulfide in water; however, the mediation mechanisms are not fully understood. In this study, we observed for the first time that the degradation of azo dyes by sulfide initially underwent a lag phase followed by a fast degradation phase. Interestingly, the presence of PCM only reduced the lag phase length of the azo dye decolorization but did not significantly enhance the reaction rate in the fast degradation phase. An analysis of the azo dye reduction and polysulfide formation indicated that PCM facilitated the transformation of sulfide into polysulfides, including disulfide and trisulfide, resulting in fast azo dye reduction. Moreover, the oxygen functional groups of the PCM, especially the quinones, may play an important role in the transformation of sulfide into polysulfides by accelerating the electron transfer. The results of this study provide a better understanding of the PCM-mediated abiotic transformation of organic pollutants by sulfide in anaerobic aqueous environments.

Insulin/IGF-1 Signaling (IIS) constrains longevity by inhibiting the transcription factor FOXO. Beyond FOXO, little is known about how phosphorylation—as mediated by IIS kinases— regulates lifespan. Here, we profiled IIS-dependent phosphorylation changes in a large-scale quantitative phosphoproteomic analysis of wild-type and three IIS mutant C. elegans strains. Our state-of-the-art analysis experimentally identified more than 15,000 phosphosites, among which 448 were differentially phosphorylated in the long-lived daf-2/insulin receptor mutant. We developed a machine-learning-based tool for systematically ranking the likely functional importance of phosphosites to guide candidate selection for follow-up validation. We show that AKT-1 pT492 inhibits DAF-16/FOXO and compensates the loss of daf-2 function, that EIF-2α pS49 potently regulates protein synthesis and daf-2 longevity, and that reduced phosphorylation of multiple germline proteins (e.g., CDK-1) apparently transmits a signal representing reduced DAF-2 signaling to the soma. Finally, kinase-substrate analysis and subsequent experimental validation confirm that casein kinase 2 negatively regulates lifespan. Our new benchmark data resource and machine-learning tool enables unprecedented access to detailed functional insights for studies of longevity.

Biochar as a multi-functional material has been widely used in the fields of catalysis and pollutant removal, but its feasibility to enhance biologic nitrobenzene reduction with the mixed culture and the long-term impact on nitrobenzene removal and microbial structure are lack of a clear clarification. This study aimed at evaluating the feasibility of biochar to enhance anaerobic reduction of nitrobenzene with a mixed culture. We initially investigated the effect of the biochar addition on nitrobenzene removal in short-term experiments and then possible enhancement mechanisms for the biochar were explored. Additionally, the impacts of different key parameters on biological nitrobenzene reduction were studied in the presence of biochar. Moreover, the long-term influence of the biochar on biological nitrobenzene removal with the mixed culture was also evaluated. The results showed that the estimated pseudo first-order rate constant of nitrobenzene removal in the presence of 1.0 g L−1 biochar was four times higher than that without biochar addition. The possible enhancement mechanisms may be owing to that biochar could activate the nitrobenzene molecules and accelerate the electron transfer between microbe and nitrobenzene. Additionally, the stimulating level of the biochar on biological nitrobenzene removal remarkably varied under different conditions. Long-term experiments demonstrated that biochar could not only increase nitrobenzene removal rate but also improve the system stability, which may be related to the shift of microbial communities that the relative abundance of nitrobenzene reduction related bacteria as well as electrochemically active bacteria was enriched in biochar addition systems.

Carbon nanotubes (CNTs) could be directly used as metal-free catalysts for the reduction of nitroaromatics by sulfide in water, but their catalytic ability need a further improvement. This study evaluated the feasibility of surface modification through thermal and radiation pretreatments to enhance catalytic activity of CNTs on nitrobenzene reduction by sulfide. The results show that thermal treatment could effectively improve the catalytic behaviors of CNTs for the reduction of nitrobenzene by sulfide, where the optimum annealing temperature was 400 °C. However, plasma radiation pretreatment didn’t result in an obvious improvement of the CNTs catalytic activity. Moreover, the possible reasons have been explored and discussed in the study. Additionally, the impacts of various operational parameters on nitrobenzene reduction catalyzed by the CNTs after an optimized surface modification were also evaluated. It was found that the rate of nitrobenzene removal by sulfide was positively correlated with CNTs doses in a range of 0.3–300 mg L−1; the optimum pH was around 8.0; higher temperature and sulfide concentration facilitated the reaction; and the presence of humic acid exhibited a negative effect on nitrobenzene reduction.

The roles and regulatory mechanisms of transcriptome changes during aging are unclear. It has been proposed that the transcriptome suffers decay during aging owing to age‐associated down‐regulation of transcription factors. In this study, we characterized the role of a transcription factor DAF‐16, which is a highly conserved lifespan regulator, in the normal aging process of Caenorhabditis elegans. We found that DAF‐16 translocates into the nucleus in aged wild‐type worms and activates the expression of hundreds of genes in response to age‐associated cellular stress. Most of the age‐dependent DAF‐16 targets are different from the canonical DAF‐16 targets downstream of insulin signaling. This and other evidence suggest that activation of DAF‐16 during aging is distinct from activation of DAF‐16 due to reduced signaling from DAF‐2. Further analysis showed that it is due in part to a loss of proteostasis during aging. We also found that without daf‐16, dramatic gene expression changes occur as early as on adult day 2, indicating that DAF‐16 acts to stabilize the transcriptome during normal aging. Our results thus reveal that normal aging is not simply a process in which the gene expression program descends into chaos due to loss of regulatory activities; rather, there is active transcriptional regulation during aging.

Smog is one of the most important environmental problems in China. Scientists have attempted to explain the causes from chemistry, physics, atmosphere and other perspectives. Many meteorologists believe that meteorology is a crucial reason. In this paper we present a new multi-view approach to visual analytics of the recent smog problems in China. This approach integrates four interrelated visualizations, each specialized in a different analysis task. To reveal the relationship between smog and meteorological attributes, we design a Correlation Detection View that simultaneously visualizes the air quality change patterns across multiple cities and related meteorological attributes. Component Trend View is used to show the variation patterns of six air components, while Aggregation View can reveal the regional overall pollution situations. A case study has been conducted using the China Air Quality Observation Data and European Centre for Medium-Range Weather Forecasts re-analysis data to verify the effectiveness of the proposed approach. By visually analyzing the meteorological data of Beijing and other major cities, we have found several interesting patterns, which prove the validity of our work in identifying sources of smog.

Anodic N doping is an effective way to improve power generation of bioelectrochemical systems (BESs), but the role of various active N dopant states of the anode on BES performance is still unclear. Herein, the effect of anodic active N dopant states on bioelectricity generation of Shewanella oneidensis MR-1 inoculated BESs particularly including microbial extracellular electron transfer (EET) was explored using experiments and theoretical simulations. It was found a positive linear correlation between the peak current density of BESs and pyrrolic N content of the anode, which would mainly ascribe to the enhancement of both direct electron transfer (DET) and mediated electron transfer (MET) of S. oneidensis MR-1. Morever, the molecule dynamic simulation revealed that such EET improvements of S. oneidensis MR-1 could be due to more remarkable reduction in the thermodynamic and kinetic resistances of the DET and MET processes with anodic doping of pyrrolic N compared to pyridinic N and graphitic N. This work provides a valuable guideline to design of high-performance anodes for potential BES applications.

The presence of submerged vegetation on river beds can change the water flow structure and alter the state of sediment motion. In this study, the incipient motion of sediment in the presence of submerged flexible vegetation in open channels was investigated in a laboratory experiment. The vegetation was simulated with flexible rubber cylinders arranged in parallel arrays. The effect of the vegetation density, water depth, and sediment grain size on the incipient motion was investigated. The experimental results indicate that the incipient motion velocity of sediment increases as the vegetation density decreases and the water depth and sediment grain size increase. With flexible plants, the incipient motion velocity of sediment is lower than it is without vegetation, and is larger than it is with rigid vegetation. A general incipient motion velocity equation was derived, which can be applied to both flexible and rigid vegetation conditions.

RNA editing increases transcriptome diversity through post-transcriptional insertion, deletion, or modification of certain bases on RNA molecules (Licatalosi and Darnell 2010). RNA editing has been found to play important roles, such as altering specific codons of neurotransmitters or ion channels (Higuchi et al. 1993; Burns et al. 1997), creating new splicing sites (Rueter et al. 1999), modifying miRNA seed sequences or their targeting sites (Kawahara et al. 2007a,b), and protecting pre-mRNA by competing with the RNAi pathway (Wu et al. 2011; Warf et al. 2012). RNA editing is a widespread phenomenon in all metazoans (Danecek et al. 2012). The vast majority of known RNA editing comprises adenosine (A) to inosine (I) deamination, which is catalyzed by a family of adenosine deaminases that act on RNA (ADARs) (Keegan et al. 2001). ADARs are essential in mammals, but their individual effects in A-to-I editing remain unclear. ADAR1+/− mutant mice died before embryonic day 14 with defects in the hematopoietic system (Wang et al. 2000). ADAR2−/− mice became progressively prone to seizures after P12 and died young (Higuchi et al. 2000). In invertebrates, Drosophila melanogaster has a single ADAR2-like gene called Dmel\Adar (also known as dADAR), the deletion of which causes locomotor incoordination and temperature-sensitive paralysis (Palladino et al. 2000). Caenorhabditis elegans has two ADAR genes, adr-1 and adr-2, but their individual roles in A-to-I editing remain unclear. Worms lacking either or both genes are viable and exhibit only minor chemotaxis defects (Tonkin et al. 2002), which makes C. elegans an excellent model to study the effects of ADARs. Next-generation sequencing and bioinformatic technologies have recently been used to identify millions of RNA editing events at the genome scale in humans (Peng et al. 2012; Ramaswami et al. 2012, 2013; Bazak et al. 2014), mice (Danecek et al. 2012; Gu et al. 2012), and fruit flies (Graveley et al. 2011). However, the RNA editome in C. elegans remains largely unknown. Thus far, only 10 genes in C. elegans had been found to be edited in noncoding regions (Morse and Bass 1999; Morse et al. 2002). It is also unclear whether RNA editing is regulated across different developmental stages. In this study, we profiled the RNA editomes of different developmental stages of the Bristol N2 wild type and three ADAR mutants using strand-specific RNA sequencing (RNA-seq) and whole-genome DNA resequencing. We also developed a new computational pipeline using a bisulfite-seq-mapping-like approach that achieved higher sensitivity and specificity. This addressed the computational challenges in identifying editing sites, including mapping reads, anchoring junction sites, and differentiating sequencing errors from true editing events (Bass et al. 2012; Piskol et al. 2013).

Postzygotic single-nucleotide mutations (pSNMs) have been studied in cancer and a few other overgrowth human disorders at whole-genome scale and found to play critical roles. However, in clinically unremarkable individuals, pSNMs have never been identified at whole-genome scale largely due to technical difficulties and lack of matched control tissue samples, and thus the genome-wide characteristics of pSNMs remain unknown. We developed a new Bayesian-based mosaic genotyper and a series of effective error filters, using which we were able to identify 17 SNM sites from ∼80× whole-genome sequencing of peripheral blood DNAs from three clinically unremarkable adults. The pSNMs were thoroughly validated using pyrosequencing, Sanger sequencing of individual cloned fragments, and multiplex ligation-dependent probe amplification. The mutant allele fraction ranged from 5%-31%. We found that C→T and C→A were the predominant types of postzygotic mutations, similar to the somatic mutation profile in tumor tissues. Simulation data showed that the overall mutation rate was an order of magnitude lower than that in cancer. We detected varied allele fractions of the pSNMs among multiple samples obtained from the same individuals, including blood, saliva, hair follicle, buccal mucosa, urine, and semen samples, indicating that pSNMs could affect multiple sources of somatic cells as well as germ cells. Two of the adults have children who were diagnosed with Dravet syndrome. We identified two non-synonymous pSNMs in SCN1A, a causal gene for Dravet syndrome, from these two unrelated adults and found that the mutant alleles were transmitted to their children, highlighting the clinical importance of detecting pSNMs in genetic counseling.

The red turpentine beetle (RTB), Dendroctonus valens LeConte (Coleoptera: Curculionidae), which is native to North America, is an invasive wood-boring insect species in China with disastrous consequences. Plant quarantine is an important method to hinder the invasion of this species. In order to distinguish RTB from the other four closely related bark beetles, Scolytus schevyrewi Semenov, Xyleborus artecomans Schedl, Xyleborus interjectus Blandford and Blastophagus minor Hartig, which are frequently intercepted at Chinese ports, we developed a nested polymerase chain reaction (PCR) assay. The specificity of the nested PCR primers to recognize RTB was confirmed by the failure to amplify genomic DNA from the other four Scolytinae species. The sensitivity was approximately 40 pg of a recombinant RTB plasmid. We evaluated the validity of the assay by analyzing bark beetles intercepted at quarantine stations. Our findings demonstrate that this specific molecular method allows for the rapid identificat...

Mutations in 23S rRNA gene are known to be associated with macrolide resistance in Mycoplasma pneumoniae (M. pneumoniae). However, these mutations alone do not fully explain the high resistance rates in Asia. The aim of this study was to investigate other possible mutations involved in macrolide resistance in M. pneumoniae.The whole genomes of 10 clinical isolates of M. pneumoniae with macrolide resistance were sequenced by Illumina HiSeq2000 platform. The role of the macrolide-specific efflux transporter was assessed by efflux-pump inhibition assays with reserpine and carbonyl cyanide m-chlorophenyl-hydrazone (CCCP).A total of 56 single nucleotide polymorphisms (SNPs) were identified in 10 clinical isolates in comparison to the reference strains M129 and FH. Strikingly, 4 of 30 SNPs causing non-synonymous mutations were clustered in macrolide-specific efflux system gene macB encoding macrolide-specific efflux pump protein of the ATP-binding cassette transporter family. In assays of the minimal inhibitory concentrations (MIC) of macrolide antibiotics in the presence of the efflux pump inhibitors caused a significant decrease of MICs, even under detectable levels in some strains.Our study suggests that macrolide efflux pump may contribute to macrolide resistance in M. pneumoniae in addition to the common point mutations in 23S rRNA gene.

How reduced graphene oxide (RGO) mediates the reductive transformation of nitroaromatic pollutants by mixed cultures and the role of its surface characteristics were evaluated in this study. Different electron donors were applied to investigate the interaction between RGO and anaerobic microbes. Moreover, the influence of the surface properties of RGO on biological nitroaromatic removal was further elucidated. The results show that RGO could achieve an approximate one-fold rate increase of nitrobenzene reduction by mixed culture with glucose as an electron donor. Selective elimination of oxygen moieties on the RGO surface, such as quinone groups, decreased the nitrobenzene transformation rate, whereas doping nitrogen into the RGO framework exhibited a positive effect. The study indicates that graphene-based carbon nanomaterials have the potential to accelerate the biological transformation of nitroaromatic compounds and that the functionalization of these carbon nanomaterials, especially through surface modification, would further enhance the conversion efficiency of contaminants.

Ce-containing Layered Double Hydroxides (Ce-LDHs) were prepared by conventional and microwave-assisted crystallization method, which were denoted as Ce-LDHs-A and Ce-LDHs-B, respectively. The dispersibility of Ce-LDHs-B is much better than that of Ce-LDHs-A; the particles of Ce-LDHs-B with particle size ranging from 20 to 70 nm are smaller than that of Ce-LDHs-A with sizes around 50-300 nm. The effect of the cerium to aluminum molar ratios, microwave crystallization time and reaction temperature on the structure of Ce-LDHs was also investigated. The results show the excessive Ce into LDHs causes distortion of the samdwich, the damage of the LDHs structure becomes more and more serious with increasing the proportion of Ce/Al. The structure of Ce-LDHs becomes more and more perfect with the increasing of the microwave crystallization time and reaction temperature.

For the solution cantilever roadheader in the construction process, the driver working strength is big, the tunnel cross section forming quality was bad, has designed the cantilever rodheader memory cutting visualization system. This system examines cantilever's position condition, the state of motion and the roadheader fuselage inclination angle through the sensor, carries on the data acquisition and processing using PLC, and through with the touch screen, the upper computer's correspondence, realizes roadheader memory cutting visualization, raised roadheader's automated level, reduced operator's labor intensity enormously, raised the tunneling efficiency obviously.

Mycoplasma pneumoniae (M. pneumoniae) is one of the common pathogens causing atypical pneumonia. In recent years, resistance to macrolides has become more common, especially in China. Previous studies have confirmed that the mutation at position 2063 in domain V of the 23S rRNA is the most prevalent, followed by the mutation at position 2064. Reported molecular detection methods for the identification of these mutations include direct sequencing, restriction fragment length polymorphism analysis, real-time polymerase chain reaction (PCR) with high-resolution melt analysis, and nested PCR-linked with capillary electrophoresis, etc. The most commonly used method for monitoring resistance-conferring mutations in M. pneumoniae is direct DNA sequencing of PCR or nested PCR products. However, these methods are time-consuming, labor-intensive or need expensive equipments. Therefore the development of rapid and sensitive methods is very important for monitoring the resistance globally.In this study, we reported a fast and cost-effective method for detecting 2063 and/or 2064 macrolide resistant mutations from specimens using a modified allele-specific PCR analysis, and all results were compared with the sequencing data. We also analyzed the clinical courses of these samples to confirm the modified allele-specific PCR results.Among 97 M. pneumoniae specimens, 88 were found to possess mutations by this method, and all modified allele-specific PCR analysis results were consistent with the sequencing data. The data of the clinical courses of these 97 cases showed that they suffered from severe pneumonia. Erythromycin showed better efficacy on cases from which no macrolide resistance mutation was found on their specimens. However, in some cases from which mutations were detected, erythromycin monotherapy had poor efficacy, and on these patients severe symptoms improved only when azithromycin was added to the treatment.The drug-resistant M. pneumoniae is very common in Beijing, China. Our modified allele-specific PCR analysis can identify erythromycin resistant mutations more rapidly from specimens than any other method currently available. Erythromycin is still effective for treating patients infected with the mutation negative M. pneumoniae, but this treatment fails to work on mutant organisms. This method can facilitate clinicians in selecting appropriate therapy within short timescales.